Introduction to Membrane Protein Antibody Discovery and Humanized Transgenic Mice

Immunogenicity is an important limitation in the development of monoclonal antibodies as therapeutic agents. Initially, attempts were made to address this issue by humanizing antibodies, but the process requires time and cost especially because of the potential compromise of the original antibody properties. Therefore, the development of fully humanized antibodies is of particular importance. The development of fully humanized antibodies using humanized transgenic mice is an effective solution. Creative Biolabs provides membrane protein immunogen preparation services and different humanized transgenic mice to support the development of fully humanized membrane protein antibodies.

What is Humanized Transgenic Mice?

Humanized transgenic mice are mice that carry functional human genes and are utilized in biomedical research and the development of clinical treatment protocols. Based on humanized transgenic mice, fully humanized antibodies can be developed. There are four commonly used techniques for obtaining humanized transgenic mice: prokaryotic microinjection, viral vector method, sperm-mediated method, and yeast artificial chromosome method.

The prokaryotic microinjection method is more costly and complicated, and the introduced exogenous genes are randomly integrated into the genome, which may lead to the mutation of endogenous functional genes or activation of oncogenes. Additionally, the number of exogenous gene integrations into the genome cannot be regulated, limiting the achievement of specific target gene integration. Furthermore, gene knockout studies cannot be conducted using this method.

The viral vector method typically includes retroviral vectors and lentiviral vectors. The retroviral vector method does not require rearrangement and allows for the integration of a single copy of the transferred gene at integration point. Embryos are placed in highly concentrated viral containers and can be co-cultured with infected cells in vitro or microinjected into chicken embryo discs. The rate of embryos integrating retroviral DNA is high. However, retroviral vectors have the disadvantage of having a narrow range of hosts for infection and a high probability of producing chimeric animals. In contrast, the lentiviral vector method has a wide range of hosts that can be infected and target nearly all types of cells. Generally, this method can infect over 90% of cells and exhibits strong genomic integration ability. However, the lentivirus itself carries potential oncogenicity, and safety improvements are needed. Additionally, the viral vector capacity is limited, allowing for the insertion of only exogenous genes smaller than 10 kb at present.

The sperm-mediated method is simple and easy to perform, relying on the natural fertilization process without damaging to the progenitor nucleus. It achieves a transgenic efficiency of up to 30% and stably integrates exogenous genes into germ cells, resulting in transgenic strains.

The yeast artificial chromosome method yields antibodies with high affinity. However, it is limited in terms of the types of antibodies produced due to the complex process of homologous recombination involving numerous Ig-adjacent gene fragments. Moreover, cloning the yeast artificial chromosome containing all Ig constant region genes poses challenge.

Membrane Protein Antibody and Humanized Transgenic Mice

The preparation of fully human membrane protein antibodies using humanized transgenic mice involves several steps. First, humanized transgenic mice are obtained. Then, the transgenic mice are immunized with the target immunogen, which prompts the mice to express the corresponding membrane protein antibodies in vivo. Compared to other techniques for obtaining human-derived membrane protein antibodies, the preparation of fully human-derived membrane protein antibodies using humanized transgenic mice offers significant advantages. On one hand, the murine system of antigen recognition and antibody mobilization remains intact in these mice, enabling them to readily recognize the human protein as a foreign substance. On the other hand, since the antibodies are produced in vivo and undergo a normal assembly and maturation process, the end product guarantees a high target binding affinity. In addition, the production of monoclonal antibody (mAb) drug candidates using transgenic mice is shorter and less expensive.

At Creative Biolabs, we are eager to share key points and experiences in membrane protein antibody discovery and humanized transgenic mice. Our aim is to assist you in smoothly running your project and achieving successful outcomes.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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